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Likaroloana tsa porous silica li ne li lokiselitsoe ke mokhoa oa sol-gel o nang le liphetoho tse ling ho fumana likaroloana tsa macroporous. s (100 × 1.8 mm id) li ne li tletse ka ho paka ka slurry. E ile ea hlahlojoa PMP kholomo karohano ea motsoako oa peptide e nang le li-peptide tse hlano (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arnkeg, leucineps ea li-album tsa leucineps leucineps leucineps leucineps ea motho ea lekang) .Tlas'a maemo a holimo a ho itlhahisa, palo ea poleiti ea theoretical ea motsoako oa peptide e phahame ho feta lipoleiti tsa 280,000 / m². Ho bapisa ts'ebetso ea karohano ea kholomo e tsoetseng pele le kholomo ea khoebo ea Ascentis Express RP-Amide, ho ile ha hlokomeloa hore ts'ebetso ea karohano ea karolo ea PMP e ne e phahametse kholomo ea khoebo mabapi le katleho ea ho arohana.
Lilemong tsa morao tjena, indasteri ea biopharmaceutical e fetohile 'maraka o ntseng o hola oa lefats'e ka keketseho e kholo ea karolo ea mmaraka. maro a 'mele, li-tissue le lisele ke mosebetsi o boima haholo ka lebaka la palo e kholo ea mefuta e ka bonoang ka har'a sampuli e le 'ngoe.Le hoja boima ba spectrometry ke sesebelisoa se sebetsang sa peptide le protheine sequencing, haeba lisampole tse joalo li kenngoa ka har'a spectrometer ea boima ka nako e le' ngoe, karohano e ke ke ea e-ba ntle. spectrometer ka nako e fanoeng4,5,6.Ho phaella moo, nakong ea karohano ea mokelikeli, li-analyte li ka tsepamisoa libakeng tse moqotetsane, kahoo li tsepamisa maikutlo analyte tsena le ho ntlafatsa MS sensitivity sensitivity.Liquid chromatography (LC) e tsoetse pele haholo lilemong tse leshome tse fetileng 'me e fetohile mokhoa o tummeng oa tlhahlobo ea proteomic7,8,9,10.
Reversed-phase liquid chromatography (RP-LC) e sebelisoa haholo bakeng sa tlhoekiso le karohano ea metsoako ea peptide e sebelisang octadecyl-modified silica (ODS) e le phase11,12,13. e hlokehang ho hlahloba li-peptide le liprotheine tse nang le likarolo tsa polar le tse se nang polar ho sebelisana le ho boloka li-analyte tsena16.Mixed-mode chromatography, e fanang ka litšebelisano tse ngata, e ka ba mokhoa o mong oa RP-LC bakeng sa karohano ea li-peptide, liprotheine, le metsoako e meng e rarahaneng.Mekhahlelo e mengata e tsoakiloeng ea mefuta e mengata e kopantsoeng1 e lokiselitsoe likarolo tse 1 tsa protheine, li-protein tse arohaneng le likarolo tse 7 tsa protheine li 'nile tsa lokisoa. 8,19,20,21.Mekhahlelo e emeng e fapaneng (WAX / RPLC, HILIC / RPLC, polar intercalation / RPLC) e loketse ho arohana ha peptide le protheine ka lebaka la ho ba teng ha lihlopha tse peli tsa polar le tse seng tsa polar22,23,24,25,26,27,28 .Ka mokhoa o ts'oanang le li-polar intercalation tsa polar le li-station tse ntle tsa polar. selectivity bakeng sa li-analyte tsa polar le tse seng tsa polar, kaha karohano e itšetlehile ka ho sebelisana pakeng tsa analyte le mohato o emeng.Litšebelisano tse ngata 29, 30, 31, 32. Haufinyane tjena, Zhang et al.30 e lokiselitse karolo e emisang ea dodecyl e felisitsoeng ea polyamine 'me e atlehile ho arola li-hydrocarbon, li-antidepressants, flavonoids, nucleosides, estrogens, le li-analyte tse ling tse ngata.Polar intercalator e na le lihlopha tse peli tsa polar le tse seng tsa polar, kahoo e ka sebelisoa ho arola li-peptide le liprotheine tse nang le hydrophobic le hydrophilicembed hydrophilicembed colummode 1. columns) li fumaneha khoebong tlas'a litšiea tsa khoebo tsa Ascentis Express RP-Amide, empa litšiea tsena li sebelisetsoa tlhahlobo ea amine 33 feela.
Phuputsong ea morao-rao, polar-embedded stationary phase (N-phenylmaleimide-embedded polystyrene) e ile ea lokisoa le ho hlahlojoa bakeng sa ho arohana ha li-peptide le li-trypsin digests tsa HSA.Mokhahlelo oa ho emisa o ile oa lokisoa ho sebelisoa leano le latelang.Likaroloana tsa silika tsa porous li ne li lokiselitsoe ho latela mokhoa o fanoeng khatisong ea rona e fetileng ka liphetoho tse ling tsa T. cetic acid e ile ea fetoloa ho lokisa likaroloana tsa silika tse nang le boholo bo boholo ba pore. Ea bobeli, ligand e ncha, phenylmaleimide-methyl vinyl isocyanate, e ile ea etsoa 'me ea sebelisoa ho hlahisa likaroloana tsa silica ho lokisetsa polar embedded stationary phase.The sephetho sa stationary phase was packed into a stainless steel column . ho sisinyeha ha mochini ho etsa bonnete ba hore bethe e ts'oanang e entsoe ka har'a kholomo.Hlahloba karohano ea kholomo e pakiloeng ea metsoako ea peptide e nang le li-peptide tse hlano;(Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) le Trypsin digest ea human serum albumin (HAS) .Motsoako oa peptide le trypsin digest ea HSA li ile tsa hlokomeloa hore li arohane ka tharollo e ntle le katleho ea kholomo ea AMP. li-peptide le liprotheine li ile tsa bonoa li rarollotsoe hantle ebile li sebetsa hantle ho kholumo ea PMP, e neng e sebetsa hantle ho feta kholomo ea Ascentis Express RP-Amide.
PEG (Polyethylene Glycol), Urea, Acetic Acid, Trimethoxy Orthosilicate (TMOS), Trimethyl Chlorosilane (TMCS), Trypsin, Human Serum Albumin (HSA), Ammonium Chloride, Urea, Hexane Methyldisilazane (HMDS), Methacryloyl Chloride, BLOYLLOYL Chloride, PO-TEMH oxide, PO-TEMH oxide, PO-TEMH HPLC Grade Acetonitrile (ACN), Methanol, 2-Propanol, le Acetone Reka ho tloha Sigma-Aldrich (St. Louis, MO, USA).
Motsoako oa urea (8 g), polyethylene glycol (8 g), le 8 ml ea 0.01 N acetic acid e ile ea tsosoa ka metsotso e 10, 'me joale 24 mL ea TMOS e ile ea eketsoa moo tlas'a maemo a batang a leqhoa. 70 ° C bakeng sa lihora tsa 12. Boima bo bonolo bo omisitsoeng bo ne bo le fatše bo boreleli ka ontong 'me bo calcined ka 550 ° C bakeng sa lihora tsa 12. Lihlopha tse tharo li ne li lokiselitsoe 'me li khetholloa ho hlahloba ho ikatisa ka boholo ba likaroloana, boholo ba pore le sebaka se ka holim'a metsi.
Ka ho fetola holim'a likaroloana tsa silica tse nang le ligand tse entsoeng pele ho li-phenylmaleimide-methylvinylisocyanate (PCMP) tse lateloang ke radial polymerization le styrene, motsoako o nang le sehlopha sa polar o ile oa lokisoa.Mokhahlelo oa ho emisa bakeng sa li-aggregates le liketane tsa polystyrene.Ts'ebetso ea ho lokisetsa e hlalositsoe ka tlase.
N-phenylmaleimide (200 mg) le methyl vinyl isocyanate (100 mg) li ile tsa qhibiliha ka toluene e omileng, 'me 0.1 mL ea 2,2'-azoisobutyronitrile (AIBN) e ile ea kenngoa ka har'a flask ea reaction ho lokisa phenylmaleimide-methyl vinyl isocyanate copolymer ka lihora tse omisitsoeng tsa C, C 6 ° ka lihora tse 3. ontong ka 40 ° C bakeng sa lihora tse 3.
Likaroloana tse omisitsoeng tsa silica (2 g) li ne li qhalakantsoeng ka toluene e omileng (100 mL), e tsosoa le ho sonicated ka 500 mL e pota-potileng ka tlas'a botlolo bakeng sa 10 min.PMCP (10 mg) e ile ea qhibiliha ka toluene 'me ea eketsoa ka dropwise ho botlolo ea ho arabela ka funnel e theoha. lihora.Joale, likaroloana tsa silika tse kopantsoeng le PMCP (100 g) li ile tsa qhibiliha ka toluene (200 ml) le 4-hydroxy-TEMPO (2 mL) e ile ea eketsoa ka pel'a 100 µL ea dibutyltin dilaurate e le catalyst.Motsoako o ile oa tsosoa ka 50 ° C mme o omisitsoe ka lihora tsa 8 °, 0.
Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), le TEMPO-PMCP-attached silica particles (1.5 g) li ile tsa qhalakanngoa ka toluene le ho hloekisoa ka naetrojene.Polymerization ea styrene e ile ea etsoa ka 100 ° C bakeng sa lihora tse 12. Sehlahisoa se hlahisoang se ile sa hlatsuoa ka methanol ka kakaretso ea 60 ° C bosiu bo omisitsoe.
Mehlala e ile ea tlosoa ka 393 K bakeng sa hora ea 1 ho fumana khatello e setseng e ka tlaase ho 10-3 Torr. Palo ea N2 e adsorbed ka khatello e lekanyelitsoeng ea P / P0 = 0.99 e ne e sebelisetsoa ho fumana palo ea kakaretso ea pore. The morphology ea likaroloana tsa silika tse se nang letho le tsa ligand-bonded, sampuli e phahameng ea electron Technologies ea Japane e ile ea hlahlojoa ka microscopy Tokyo. (silica e se nang letho le likaroloana tsa silika tse entsoeng ka ligand) li ne li behoa holim'a mohala oa aluminium ho sebelisa sekhomaretsi sa carbon tape. Khauta e ne e pentiloe holim'a mehlala e sebelisang Q150T sputter coater, 'me 5 nm Au layer e ne e behiloe holim'a mehlala.Sena se ntlafatsa ts'ebetso ea ts'ebetso ka ho sebelisa li-voltage tse fokolang' me se fana ka lijo-thollo tse ntle, tse batang tsa sputtering, EA2 Eleron ea Amerika, EA2 Eleron e ne e le sputtering e batang ea USA1. e sebelisetsoang tlhahlobo ea motheo.A Malvern (Worcestershire, UK) Mastersizer 2000 particle size analyzer e ne e sebelisetsoa ho fumana kabo ea boholo ba likaroloana.Likaroloana tsa silica tse hlobotseng le li-silica particles tsa ligand-bonded (5 mg ka 'ngoe) li ile tsa qhalakanngoa ka 5 mL ea isopropanol, sonicated bakeng sa 10 min, vortextical benched on the Master of the 5. e etsoa ka sekhahla sa 5 °C ka motsotso ho feta mocheso oa 30 ho isa ho 800 °C.
Litšepe tse sa hloekang tse entsoeng ka khalase tse nang le litšepe tse tšesaane tsa litekanyo (id ea 100 × 1.8 mm) li ne li pakiloe ho sebelisoa mokhoa oa ho paka seretse, ho sebelisoa mokhoa o tšoanang o sebelisitsoeng ho Ref.31.Kolumo ea tšepe e sa hloekang (e kentsoeng ka khalase, 100 × 1.8 mm id) e nang le "outlet fitting" e nang le 1 µm frit e ne e hokahantsoe le sephutheloana sa slurry packer (Alltech Deerfield, IL, USA). Lokisetsa slurry ea mokhahlelo o emeng ka ho emisa 150 mg ea mokhahlelo o emisang oa menotha o sebelisitsoeng ho menotha ho 1. slurry solvent hammoho le propelling solvent.Tlatsa kholumo ka tatellano ka ho sebelisa likhatello tsa 100 MP bakeng sa metsotso ea 10, 80 MP bakeng sa metsotso ea 15, le 60 MP bakeng sa metsotso e 30. Nakong ea ho paka, ho sisinyeha ha mechine ho ne ho sebelisoa ka li-shaker tse peli tsa GC (Alltech, Deerfield, IL e pakelang junifomo ea IL, USA) ho etsa bonnete ba hore ho na le ts'enyehelo ea li-column tse thibelang li-packers. kholumo.Hakolla kholomo ho tloha yuniti ea ho paka e slurry ebe o hokela e 'ngoe e loketseng ho kena le ho sistimi ea LC ho lekola ts'ebetso ea eona.
LC pump (10AD Shimadzu, Japan), injector (Valco (USA) C14 W.05) e nang le 50nL injection loop, membrane degasser (Shimadzu DGU-14A), UV-VIS capillary window e hahiloe Special µLC device detector (UV-2075) le khalase e nang le li-microcolum tse nyenyane haholo tse kopanyang likhalase tse nyenyane haholo. Ening. Kamora ho paka, li-capillaries (50 μm id 365 le ho fokotsa li-capillaries tsa bonngoe (50 μm) li ile tsa kengoa sebakeng sa 1/16 ″ sa kopano e fokotsang. Northampton, MA).
Albumin ho tloha serum ea motho, phofo ea lyophilized, ≥ 96% (agarose gel electrophoresis) 3 mg e kopantsoeng le trypsin (1.5 mg), 4.0 M urea (1 mL), le 0.2 M ammonium bicarbonate (1 mL) . Tharollo e ile ea tsosoa ka metsotso e 10 'me e bolokoa ka bateng ea metsi ka lihora tse 60 tsa T1. Sefa tharollo 'me u boloke ka tlase ho 4 °C.
Karohano ea metsoako ea peptide le tšilo ea li-trypsin ea HSA e ile ea hlahlojoa ka thoko ho likholomo tsa PMP. Hlahloba karohano ea motsoako oa peptide le trypsin digest ea HSA ka kholomo ea PMP 'me u bapise liphetho le kholomo ea Ascentis Express RP-Amide. Nomoro ea poleiti ea theory e baloa ka tsela e latelang:
Litšoantšo tsa SEM tsa likaroloana tsa silika tse se nang letho le likaroloana tsa silica tse kopantsoeng le ligand li bontšoa ho FIG.2 .sem setšoantšo sa likaroloana tsa silio (a, b) bontša hore, karolo ea Sillica e sa tloaelehang.
Ho hlahloba litšoantšo tsa electron microscope ea likaroloana tse se nang letho tsa silica (A, B) le likaroloana tsa silica tse kopantsoeng le ligand (C, D).
Likarolo tsa boholo ba likaroloana tsa silika tse se nang letho le likaroloana tsa silika tsa ligand-bonded li bontšoa ho Setšoantšo sa 3 (A) . Volume-based particle size distribution curves e bontšitse hore boholo ba likaroloana tsa silika bo ile ba eketseha ka mor'a phetoho ea lik'hemik'hale (setšoantšo sa 3A) . 3.36 μm, ha e bapisoa le thuto ea rona e fetileng e nang le papatso (0.5) boleng ba 3.05 μm (polystyrene-bound silica particles) 34. Sehlopha sena se ne se e-na le kabo e fokolang ea likaroloana ha e bapisoa le thuto ea rona e fetileng ka lebaka la likarolo tse fapaneng tsa PEG, urea, TMOS, le acetic acid ka karabelo ea motsoako o moholo oa Prene MP. .Sena se bolela hore ts'ebetso ea holim'a silika e nang le styrene e kentse feela lesela la polystyrene (0.97 µm) holim'a silica, athe karolong ea PMP botenya ba lera e ne e le 1.38 µm.
Kabo ea boholo ba likaroloana (A) le kabo ea boholo ba pore (B) ea likaroloana tsa silika tse se nang letho le likaroloana tsa silika tse tlamang ligand.
Boholo ba pore, bophahamo ba pore le sebaka sa holim'a likaroloana tsa silika tsa thuto ea morao-rao li fanoe ho Lethathamo la 1 (B) . Litlhaloso tsa PSD tsa likaroloana tsa silika tse se nang letho le likaroloana tsa silika tse entsoeng ka ligand li bontšoa ho Setšoantšo sa 3 (B) . Liphetho li bapisoa le thuto ea rona e fetileng. boholo bo fokotseha ka 69 ka mor'a phetoho ea lik'hemik'hale, joalokaha ho bontšitsoe Lethathamong la 1 (B), 'me phetoho ea lekhalo e bontšoa ho Setšoantšo sa 3 (B) .Ka mokhoa o ts'oanang, palo ea pore ea likaroloana tsa silika e fokotsehile ho tloha 0.67 ho 0.58 cm3 / g ka mor'a phetoho ea lik'hemik'hale. . Joalokaha ho bontšitsoe Lethathamong la 1 (B), sebaka se ka holimo (m2 / g) sa likaroloana tsa silika le sona se fokotsehile ho tloha ho 116 m2 / g ho ea ho 105 m2 / g ka mor'a phetoho ea lik'hemik'hale.
Liphetho tsa tlhahlobo ea mantlha ea mohato o emeng li bonts'itsoe ho Lethathamo la 2. The carbon loading ea boemo ba hona joale ba ho emisa ke 6.35%, e leng tlase ho feta carbon loading ea thuto ea rona e fetileng (polystyrene bonded silica particles, 7.93% 35 le 10.21%, ka ho latellana) 42. The carbon loading of the current phase, SP , SP , SP, SP, 35% le 10.21%. lihlahisoa tse kang phenylmaleimide-methylvinylisocyanate (PCMP) le 4-hydroxy-TEMPO li ile tsa sebelisoa.Palo ea boima ba naetrojene ea karolo ea hona joale e emeng ke 2.21%, ha e bapisoa le 0.1735 le 0.85% ka boima ba naetrojene lithutong tse fetileng, ka ho latellana.Sena se bolela hore wt % ea nitrogen loadary. 4) le (5) e ne e le 2.7% le 2.9%, ka ho latellana, ha carbon loading ea sehlahisoa sa ho qetela (6) e ne e le 6.35%, joalokaha ho bontšoa ho Lethathamo la 2. Ho lahleheloa ke boima ba 'mele ho ile ha hlahlojoa le karolo ea PMP stationary,' me lekhalo la TGA le bontšoa ho Setšoantšo sa 4. Lekhalo la TGA le bontša tahlehelo ea boima ba 'mele ea 3 e nang le k'habone e nang le 8.6% feela e nang le li-liga tsa 8.6 ka lebaka la ho lahleheloa ke boima ba lik'hemik'hale tsa 6% feela. hape N, O, le H.
Phenylmaleimide-methylvinylisocyanate ligand e khethiloe bakeng sa phetoho ea holim'a likaroloana tsa silica hobane e na le lihlopha tsa polar phenylmaleimide le lihlopha tsa vinylisocyanate.Lihlopha tsa vinylisocyanate li ka tsoela pele ho itšoara ka styrene ka ho phela ka radical polymerization. mide moiety ha e na tefiso ea sebele ka pH e tloaelehileng. Polarity ea mohato o emeng o ka laoloa ke tekanyo e nepahetseng ea styrene le nako ea ho itšoara ka polymerization ea mahala. Mohato oa ho qetela oa karabelo (free-radical polymerization) ke oa bohlokoa haholo 'me o ka fetola polarity ea mohato o tsitsitseng. ka tsela e fapaneng.SPs e lokiselitsoeng ka mekhahlelo e fapaneng ea styrene e na le mefuta e sa tšoaneng ea carbon loadings.Hape, laela mekhahlelo ena e emeng ka har'a litšiea tsa tšepe e sa hloekang 'me u hlahlobe ts'ebetso ea tsona ea chromatographic (selectivity, resolution, N value, joalo-joalo) .Ho itšetlehile ka liteko tsena, ho ile ha khethoa mokhoa o optimized ho lokisetsa mohato oa ho emisa PMP ho etsa bonnete ba hore polarity e laoloang le ho hlahloba hantle.
Metsoako e mehlano ea peptide (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) le eona e ile ea hlahlojoa ho sebelisoa mohala oa PMP o sebelisang mohato oa selefouno;60/40 (v/v) acetonitrile/metsi (0.1% TFA) ka sekhahla sa phallo ea 80 μL/min. Tlas'a maemo a ntlafetseng a ho hlakisa, nomoro ea poleiti ea theoretical (N) ka kholomo (100 × 1.8 mm id) ke 20,000 ± 100 (lipoleiti tse tharo tsa 3MP bakeng sa kholomo ea 3MP e fana ka 0). li-chromatogram li bonts'itsoe ho Setšoantšo sa 5A. Tlhahlobo e potlakileng ho kholumo ea PMP ka sekhahla se phallo se phahameng (700 μL/min), li-peptide tse hlano li ile tsa hlakoloa nakong ea motsotso o le mong, boleng ba N bo ne bo le ntle haholo, 13,500 ± 330 kholomong (100 × 1.8 mm id), E tsamaellana le 135 × 1. (100 × 1.8 mm id) li ne li tletse lotho tse tharo tse fapaneng tsa PMP stationary phase to check reproducibility.The analyte concentration for each column e ngotsoe ho sebelisoa maemo a holimo a elution le palo ea lipoleiti tsa khopolo-taba N le nako ea ho boloka ho arola motsoako o tšoanang oa teko kholumong e 'ngoe le e 'ngoe. The reproducibility columns for the PMPble columns with the PMPble columns wellly displayed in the PMPble columns are very shown. Boleng ba RSD, joalo ka ha ho bonts'itsoe ho Lethathamo la 3.
Karohano ea motsoako oa peptide holim'a kholomo ea PMP (B) le kholomo ea Ascentis Express RP-Amide (A);mobile phase 60/40 ACN/H2O (TFA 0.1%), PMP columna litekanyo (100 × 1.8 mm id);analytical Taelo ea elution ea metsoako: 1 (Gly-Tyr), 2 (Gly-Leu-Tyr), 3 (Gly-Gly-Tyr-Arg), 4 (Tyr-Ile-Gly-Ser-Arg) le 5 (leucine) acid enkephalin)).
Kholomo ea PMP (ID ea 100 × 1.8 mm) e ile ea hlahlojoa bakeng sa karohano ea tšilo ea tryptic ea albumin ea serum ea motho ka chromatography ea mokelikeli o phahameng. Chromatogram e setšoantšong sa 6 e bonts'a hore sampole e arohane hantle 'me qeto e ntle haholo.Litlhaloso tsa HSA li ile tsa hlahlojoa ho sebelisoa lebelo la phallo ea 100min/totri3/mobile 100 µg le 100 min / 3 µ µ µ. 1% TFA Joalo ka ha ho bonts'itsoe ho chromatogram (setšoantšo sa 6), tšilo ea HSA e arotsoe ka litlhōrō tse 17 tse tsamaellanang le li-peptide tse 17. Ho sebetsa hantle ha karohano ea tlhoro e 'ngoe le e 'ngoe ho HSA digest ho ile ha baloa 'me boleng bo fanoa ho Lethathamo la 5.
Tryptic digest ea HSA (100 × 1.8 mm id) e ile ea aroloa holim'a kholomo ea PMP;sekhahla sa phallo (100 µL/min), karolo ea selefouno 60/40 acetonitrile/metsi ka 0.1% TFA.
moo L e leng bolelele ba kholomo, η ke viscosity ea mohato oa selefouno, ΔP ke khatello ea mokokotlo oa mokokotlo, 'me u ke lebelo la mola oa mohato oa selefouno. The permeability ea kholomo ea PMP e ne e le 2.5 × 10-14 m2, tekanyo ea phallo e ne e le 25 μL / min, le 60/40 v / v / v. e ne e tšoana le ea thuto ea rona e fetileng Ref.34.The permeability ea kholomo e tletse ka likaroloana tse ka holimo-limo ke: 1.7 × 10-15 bakeng sa 1.3 μm likaroloana, 3.1 × 10-15 bakeng sa 1.7 μm likaroloana, 5.2 × 10-14 × 2 μm 2 × 2 × 2. μm likaroloana 43. Ka lebaka leo, ho khoneha ha karolo ea PMP ho tšoana le ea 5 μm core-shell particles.
moo Wx e leng boima ba kholomo e nang le chloroform, Wy ke boima ba kholomo e nang le methanol, le ρ ke boima ba mokelikeli.Densities of methanol (ρ = 0.7866) le chloroform (ρ = 1.484).Kakaretso ea porosity ea SILICA PARTICLES 108 × 18mm CARTICLES-108 mm Ikholomo ea 1-8 × 18 × 1. Litšiea tsa Urea 31 tseo re ithutileng tsona pele e ne e le 0.63 le 0.55, ka ho latellana. Sena se bolela hore ho ba teng ha li-urea ligands ho fokotsa ho phunyeletsa ha karolo e emeng. Likarolo tse emeng tsa mofuta oa 8 li-ligand tsa C18 li khomaretsoe ho likaroloana tsa silika e le liketane tse melang, ha ka mekhahlelo ea mofuta oa polystyrene, ho thehoa A polymer layer e batlang e le teteaneng.
Setšoantšo sa 7A, B se bontša kholomo ea PMP (100 × 1.8 mm id) le kholomo ea Ascentis Express RP-Amide (100 × 1.8 mm id) ho sebelisoa maemo a tšoanang a elution (ke hore, 60/40 ACN/H2O le 0.1% TFA).) ea morero oa van Deemter.Metsoako e khethiloeng ea peptide (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) e lokisitsoe ka 20 µL/ Sekhahla sa phallo e tlase bakeng sa likholomo ka bobeli ke 800 µL/min. Bonyane ba 800 µL/min. ke Express RP-Amide kholomo e ne e le 2.6 µm le 3.9 µm, ka ho latellana. Litekanyetso tsa HETP li bontša hore katleho ea karohano ea kholumo ea PMP (100 × 1.8 mm id) e molemo haholo ho feta e fumanehang khoebong ea Ascentis Express RP-Amide kholomo (100 × 1.8 1.8 mm Id) e bonts'a phallo e ntseng e eketseha ka 100 × 1.8 mm Deemter. e seng ea bohlokoa ha e bapisoa le thuto ea rona e fetileng.Katleho e phahameng ea karohano ea kholomo ea PMP (100 × 1.8 mm id) ha e bapisoa le kholomo ea Ascentis Express RP-Amide e thehiloe ho ntlafatso ea sebopeho sa likaroloana, boholo, le mekhoa e rarahaneng ea ho paka mela e sebelisoang mosebetsing oa hona joale34.
(A) van Deemter plot (HETP khahlano le mobile phase linear velocity) e fumanoe ka ho sebelisa kholomo ea PMP (100 × 1.8 mm id) ka 60/40 ACN/H2O le 0.1% TFA. (B) van Deemter plot (HETP khahlanong le mobile phase linear velocity) e fumanoeng ho sebelisoa RP- Ascentis 1 × 1 kholomo ea Ascentis × 1 RP-Ascentis Express 0 ACN/H2O e nang le 0.1% TFA.
Polar-embedded polystyrene stationary phase e ile ea lokisoa le ho hlahlojoa bakeng sa karohano ea metsoako ea maiketsetso ea peptide le trypsin digests ea human serum albumin (HAS) in high performance liquid chromatography.The chromatographic performance of PMP columns for peptide mixes is excellent in karohano katleho le resolution.The e ntlafetseng ea karohano ea mefuta e fapaneng ea kholomo ea mabaka a joalo ka karohano ea boholo ba silika e le ka lebaka la boholo ba karohano ea silika, ka lebaka la boholo ba karohano ea silika, ka lebaka la ts'ebetso e ntlafalitsoeng ea likaroloana tsa silika, joalo ka boholo ba karohano ea silika. particles, controlled synthesis of the stationary phase, and complex column packing.Ho phaella ho katleho e phahameng ea karohano, khatello e tlaase ea mokokotlo oa mokokotlo ka litekanyetso tse phahameng tsa phallo ke molemo o mong oa phallo ena e tsitsitseng.PMP columns display good reproducibility and can be used for the analysis of peptide mixs and trypsin tshilo ya diprotheine tse fapa-fapaneng. ography.Nakong e tlang, litšiea tsa PMP li tla boela li hlahlojoe bakeng sa karohano ea liprotheine le li-antibodies tsa monoclonal.
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